Project III- Molecular biology of AAV in liver Molecular characterization of liver gene transfer and its oncogenic potential A substantial advantage of AAV for genetic diseases is its ability to confer long-term and high-level transgene expression in liver. This project will evaluate several interesting aspects of molecular biology as it relates to mechanisms of persistence and potential safety considerations. The first specific aim will address the potential for recombination between the transduced vector genome and the latent wild-type AAV that occurs subsequent to a natural infection. The second specific aim will undertake a systematic evaluation of the molecular state and structure of persistent AAV genomes with a focus on the frequency and distribution of integration sites by adapting the state-of-art massive parallel genome wide pyrosequencing technology for studying AAV integration. The final specific aim will undertake experiments to actually quantitate the oncogenic potential of AAV following the delivery to liver. These studies will be performed in two murine models for dyslipidemia including LDLR/Apobec-1 double KO and LDLR/Apobec-1/MTP triple KO mice. In addition, the consequence of chronic inflammation caused by steatohepatitis on the oncogenic potential of AAV will be studied Project III will rely on Projects I and II for obtaining tissues from nonhuman primates who have receivedAAV vectors as part of its goal to undertake molecular characterization. This project will rely on the expertise of Dr. Rader in Project II to conduct the tumors studies in the murine models for lipidemia. Lay description. This project will evaluate the molecular structure of the transferred gene as is resides in the liver cells. Animal studies will be performed to determine if the vector causes tumors. RELEVANCE (See instructions):